Effects of long-term storage on the detection of proteins, DNA, and mRNA in tissue microarray slides

J Histochem Cytochem. 2011 Dec;59(12):1113-21. doi: 10.1369/0022155411423779.

Abstract

Storage of tissue slides has been claimed to induce dramatically reduced antigen detection particularly for immunohistochemistry (IHC). With tissue microarrays, the necessity to serially cut blocks in order to obtain as much material as possible is obvious. The presumed adverse effect of storage might hamper such an approach. The authors designed an experimental setting consisting of four different storage conditions with storage time of tissue slides of up to 1 year. Detection of proteins, DNA, and mRNA was performed using IHC and in situ hybridization techniques. Slight but significant changes in IHC occurred over time. The most important factor is the primary antibody used: four showed no significant changes, whereas limited decreases in 8 antibodies could be detected by image analysis. Whether the antigen was nuclear or cytoplasmic/membranous did not matter. No major differences between different storage conditions could be shown, but storage at 4C was overall the best procedure. Furthermore, gene copy number aberrations, chromosomal translocations, and the presence of mRNA could be detected on slides stored up to 1 year. In conclusion, in tissues optimally formalin fixed and using modern histological techniques, only minute changes in tissue antigenicity are induced by long-term storage.

MeSH terms

  • DNA / analysis*
  • Factor Analysis, Statistical
  • Formaldehyde
  • Gene Dosage
  • Humans
  • Immunohistochemistry
  • In Situ Hybridization, Fluorescence
  • Keratin-7 / metabolism
  • Ki-67 Antigen / metabolism
  • Neoplasms / metabolism
  • Paraffin Embedding
  • Proteins / analysis*
  • RNA, Messenger / analysis*
  • Receptor, ErbB-2 / metabolism
  • Specimen Handling*
  • Temperature
  • Time Factors
  • Tissue Array Analysis
  • Tissue Fixation
  • Translocation, Genetic
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • Keratin-7
  • Ki-67 Antigen
  • Proteins
  • RNA, Messenger
  • Tumor Suppressor Protein p53
  • Formaldehyde
  • DNA
  • ERBB2 protein, human
  • Receptor, ErbB-2